Gene Transfer to Plants

Part of the series Springer Lab Manual pp 30-38

Introduction II Agrobacterium-rhizogenes,a Natural Transformation System

  • J. B. Bade
  • , B. Damm

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The protocol detailed below describes an efficient method for Agrobacterium tumefaciens-mediated transformation of Brassica napus. cv Westar based on two earlier published procedures (Radke et al. 1988; De Block et al. 1989). Hypocotyl explants are cocultivated in liquid medium with the nopaline-type Agrobacterium strain MOG301 carrying different binary vectors comprising the gus-intron reporter gene and a selectable marker. Transgenic plants are produced with kanamycin, hygromycin B, phosphinothricin, or chlorsulfuron as selective agents.